JP2960867B2 - Oil-in-water nanoemulsions useful as ophthalmic vehicles and their preparation - Google Patents

Abstract

The nanoemulsion comprises: 0.1-10% (w/v) of an oil; 0.1-10% (w/v) of a non-ionic surface active agent; a maximum of 0.01% (w/v) of benzalkonium chloride; a drug or a precursor or active substance; and optionally, one or several of the following components: an isotonizing agent, a viscosity modifying agent or stabilizer, a buffer and/or an antioxidant in variable proportions. The process comprises: (a) preparing an aqueous phase containing, among others, a non-ionic surface active agent; (b) preparing an organic phase containing dissolved an oil and an active substance in an organic water miscible solvent in all proportions; (c) including the organic phase to the aqueous phase under moderate agitation; and (d) totally evaporating the organic solvent and part of the water up to the final desired volume. Important applications as an ophthalmic vehicle in preparations used to treat eye disorders and diseases.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to the field of drug release by oil-in-water emulsions, especially for ophthalmic use. The present invention provides a vehicle that increases corneal penetration of an active agent contained in a composition.

[0002]

BACKGROUND OF THE INVENTION The most commonly used vehicle for the treatment of eye disorders and diseases is an aqueous solution. However, the aqueous solution applied to the eye is delacrymation (del
It is rapidly diluted and eliminated by aeration and mechanical action of blinking, and is largely lost through the nasolacrimal duct. Thus, most of the administered drug is eliminated before penetrating the cornea and sclera, reducing the bioavailability of the drug to the eye. On the other hand, many of the medicaments that can be applied to the eye are extremely lipophilic and are therefore very sparingly soluble in water, so they must be applied in the form of a suspension or ointment, and in some cases after application. May cause irritation and discomfort. Moreover, in practice, the drug is required to be dissolved so as to be absorbed before it is eliminated from the surface of the eye, and in the case of suspension, the bioavailability is reduced.

[0003] Other types of vehicles have been developed to increase the residence time of drugs at the surface of the eye. Among these, vehicles whose biocompatibility has been improved by increasing the viscosity are like hydrogels or ophthalmic ointments. In the case of hydrogels, a satisfactory improvement in the bioavailability of the drug has not been obtained. Some ophthalmic ointments are preferred for use at night because their application is very cumbersome and the eyes are blurred after application. Similarly, a great number of new vehicles such as liposomes, nanoparticles, etc. have been developed, but with considerable success in terms of bioavailability, most of them are not stable, tolerable and difficult to industrialize. There is a problem.

[0004] Different types of emulsions have been proposed as vehicles for drug release in the eye. Among these, EP-A-0 521 799 Al describes oil-in-water emulsions for the release of hydrophobic, amphoteric and lipophilic drugs. The composition includes an oil, a phospholipid and an amphoteric surfactant. Although the role of phospholipids is essential for the stability of the emulsions of this invention, phosphatidylcholine and its derivative lysophosphatidyl are essentially cataracts.
other authors have been described by others ([1] "Effects of
Lysophosphatidylcholine and phospholipase A
・ On the lens (Effects of lysop)
phosphatidyl choline and ph
ospholipase A on the len
s) ", Cotlier, E.,
Baskin, M. and Kresca, L., Investigative ophthalmology.
Ophthalmology), Vol. 14, No. 9: 697-701 (1975). (2) “Phospholipid effects on the rat lens transport systems (Phospholipid ef)
facts on the rat lens trans
Port systems ", written by Kador, P.F. and Kinoshita, J.H., Kinoshita, JH, Exp. Eye Res., 2nd ed.
6, 657-665 (1978)].

On the other hand, health authorities in most countries require the use of preservatives in ophthalmic products. This patent discloses thimerosal-chlorobutanol,
Since it is not effective if used separately, it is 0.01
It claims to be used in a concentration of ~ 0.2% (weight / volume) and in combination with benzalkonium chloride at a concentration of 0.02% (weight / volume) when used alone.

[0006] The present invention provides ophthalmic formulations in the form of nanoemulsions which are stable for long periods without the inclusion of phospholipids in the composition. On the other hand, the subject formulations of the present invention meet the European, British and US Pharmacopoeia criteria of using up to 0.01% (w / v) of benzalkonium chloride.

US Pat. No. 5,171,566 also discloses an oil-in-water emulsion which contains soybean oil and soybean lecithin as emulsifiers. In addition to lecithin, this type of emulsion also contains phosphatidylcholine, and may therefore have the same problem of toxicity described above. Similarly, they contain other stabilizers such as cholesterol or phosphatidylic acid. The emulsion may be lyophilized or kept at 4 ° C. This composition has the same disadvantages as the above patent and contains only flurbiprofen (2-fluoro-α-methyl [1,1-biphenyl] -4-acetic acid) and its esters. Unlike the present invention, it does not claim or describe the effect of improving biocompatibility, but rather limits it to mentioning the presence of the drug in the bodily fluid of the rabbit. On the other hand, we have observed that the nanoemulsions of the present invention can increase the bioavailability of pharmaceuticals in the eye by about 4-fold.

[0008] European Patent Application 0 480 690 Al describes an ophthalmic product of the emulsion type, but deals with a product which is substantially different from that of the present invention. The above-mentioned application discloses tepoxaline having a translucent to transparent appearance which is a characteristic characteristic of microemulsions having an oil droplet size of 0.005 to 0.5 μm.
Claims the preparation of microemulsions. This appearance greatly limits its industrial productivity. The nonionic surfactant used is polysorbate (polys).
and the concentration of the preservative (s) is from 0.02 to
0.7% (weight / volume). The present invention deals with different compositions because, instead of the microemulsion, it is a nanoemulsion that is neither transparent nor translucent (transmittance at 520 nm is less than 70%). Similarly, the amount of preservative used is much less than that used in the aforementioned patents where the toxicity of the preservative is a problem at this concentration.

[0009]

SUMMARY OF THE INVENTION It is an object of the present invention to improve the bioavailability by keeping low levels of preservatives, which are required by the pharmacopoeia standards of many countries but have considerable toxic effects on the eyes. Another object of the present invention is to provide a useful vehicle for ophthalmic use.

[0010] The present invention provides an oil-in-water emulsion-type preparation having high bioavailability in pharmaceutical eyes in a vehicle. This emulsion contains a cataract (cat) as in the case of potentially irritating products and lecithin in the composition.
It is stable during storage without any products that cause arcogenic processes. Also according to the formulations of the present invention, preservatives (benzalkonium chloride) in small concentrations that do not meet the Pharmacopoeia standards for ophthalmic products can be used in the other types of formulations described above. On the other hand, the emulsions of the present invention can be obtained using conventional emulsifying equipment, such as a rotary stirrer or other pressurized homogenizer.

Although most national health regulations require that preservatives be included in multidose-type ophthalmic products, in practice all preservatives used are of considerable toxicity to the eyes. There is action. Therefore, having an ophthalmic vehicle that can use preservatives at low concentrations represents a very important advantage for ophthalmic formulation safety. As described above, the present invention relates to the method of the present invention, which is performed in the range of 0.01 to 0.1.
2% (weight / volume) chlorobutanol and 0.01 to 0.1%
Claims the combined use of 2% (weight / volume) thimerosal (benzalkonium chloride is 0.02% in order to be able to interact with the surfactant or to be absorbed in the oil used) Unlike those described in European Patent Application No. 0 521 799 Al, preservatives (since weight / volume has no effect) (0.005 to 0.01%
An ophthalmic vehicle that can be used in low concentrations (weight / volume benzalkonium chloride).

[0012]

SUMMARY OF THE INVENTION In accordance with the present invention, an oil-soluble or partially oil-soluble medicament is included in an oil-in-water emulsion to be administered to the eye, thereby providing a higher concentration than other compositions. Its biological applicability is enhanced. The vehicle contains an oil and a nonionic surfactant, and sufficient preservatives to comply with pharmacopoeia standards. The oils that form part of the emulsion may be vegetable oils, animal oils, mineral oils, fatty acids, medium chain triglycerides, fatty alcohols or any combination of oils and oils that are well tolerated by the eye.

The most preferred oils are the medium chain triglycerides because of their high solubility in water, their density, their insensitivity to oxidation, and their excellent tolerance in the eyes. Among these, fractionated C _8-10 fatty acid triglycerides of coconut oil, and C _8-10
Propylene glycol esters of medium chain saturated fatty acids are excellent. Polyethylene glycol esters and glycerides should also be addressed. Olive oil, sunflower seed oil and sesame oil with acid numbers below 0.5 are worth mentioning as vegetable oils. The oil is preferably 0.1 to 10% (weight / volume) in the composition of the present invention.

Examples of nonionic surfactants are polyoxyethylene-polyoxypropylene copolymers, preferably poloxamer 188 and poloxamer 407. These surfactants are 10% (weight / weight) when administered to the eye.
Even volume) concentrations are very tolerable, are not irritating to the eyes and do not cause harm. By using these surfactants at appropriate concentrations, the nanoemulsion can be kept stable without using other co-surfactants. The preferred nonionic surfactant concentration for the composition of the present invention is 0.1 to 10% (weight / volume).

[0015] The compositions of the present invention may include other medicaments used in ophthalmology. Examples of anti-glaucoma drugs include carteolol-based, betaxolol, atenolol,
Mention may be made of carbonic anhydrase inhibitors such as timolol-based and metazolamide. Examples of antibiotics include chloramphenicol, and examples of anti-inflammatory drugs include indomethacin, piroxicam, dichlorophenacic acid, ibuprofen, dexamethasone and clobetasone. Another type of medicament is cyclosporin A, acyclobeer (2-amino-1,9-dihydro-9-[(2-hydroxy-ethoxy) methyl] -6H-purin-6-one) and acid chromogly Kate. Depending on the nature of the oil, the compositions of the present invention may include antioxidants to prevent oxidation of the oil.

The composition of the present invention comprises an isoosmotic agent such as mannitol, glycerol, sorbitol or glucose; a viscosity modifier such as hydroxypropylmethylcellulose, a polyacrylic acid derivative or sodium carboxymethylcellulose; sodium edatate (sod)
stabilizers such as sodium citrate and citric acid; buffers such as sodium and potassium phosphates, sodium citrate, sodium carbonate and sodium bicarbonate. Stabilizing the composition of the present invention by using a polyacrylic acid polymer at a certain concentration between 0.1 and 0.5% (weight / volume) and creaming by flocculation of oil droplets and phase separation. Even you can avoid. These compositions have an apparent white gel-like appearance with a certain viscosity.

As noted above, the present invention provides an oil-in-water nanoemulsion useful as an ophthalmic vehicle, comprising: (a) about 0.1-10% (weight / weight) (B) about 0.1 to 10% (w / v) nonionic surfactant; (c) about 0.01% (w / v) or less;
Benzalkonium chloride as preservative (d) 0.01-5% (w / v) of medicament, pharmaceutical precursor or biologically active substance (e) one or more of the following optional ingredients: : Various amounts of isoosmotic agent; viscosity modifier; stabilizer, buffer and / or
Or antioxidants.

The emulsion of the present invention has a transmittance measured at 520 nm of less than 70%, a pH between 5 and 8, and an osmotic pressure of 250-400 mOsm / kg.
The appearance of these emulsions is thin milky. The composition of the present invention may be sterilized by filtration as long as the oil droplets can be formed, or may be sterilized in an aqueous phase and an oil phase, and then mixed and emulsified depending on the appearance.

Contrary to what would be expected, the medicament contained in the composition of the present invention penetrates the cornea up to six times more than the same medicament in aqueous solution in an "in vitro" experimental model. . Similarly, when a composition of the present invention is infused into rabbit eyes, the amount of drug at "in vivo" levels in aqueous bodily fluids will be approximately four times higher than that obtained with an aqueous solution of the same drug. This increased bioavailability often means that there is a great advantage since the daily infusion or dosage of these medicaments used to treat chronic eye diseases can be reduced.

The compositions of the present invention may be prepared in different ways. One method involves preparing the water and oil phases separately. The aqueous phase contains the appropriate proportions of non-ionic surfactants, iso-osmotic agents, preservatives and pH buffer systems. The oil phase contains the whole and part of the active substance dissolved in the oil and can contain antioxidants. To prepare the emulsion, the oil phase is added to the aqueous phase under moderate stirring, and then a homogenizer of the Ultra-turrax type (Junk and Kunkel) until an average particle size of less than 5 μm is obtained. (Janke and F
unkel), Staofen,
Germany] to reduce the particle size. Oil droplets of this size can also be obtained using a high pressure homogenizer or any device capable of suitably reducing particle size.

According to another particular preparation method of the invention, the usual exposure of the product to a temperature of 45-85 ° C., which affects the heat-sensitive active principle or affects the oxidation and decomposition reactions of the components of the formulation. In contrast to the above temperature, an oil-in-water emulsion having an average oil droplet size of 200 nm can be obtained at a temperature of 35 ° C. or lower.

This preparation method comprises preparing an aqueous phase in the same manner as described above and preparing an organic phase containing the oil and an active substance which is completely or partially soluble in the oil, wherein both are acetone , Soluble in water such as ethanol or tetrahydrofuran and soluble in organic solvents having a dielectric constant greater than 15. The above organic phase is added to the aqueous phase under moderate stirring, and a part of the organic solvent and water is removed under reduced pressure by an appropriate evaporation system.
Removed at a temperature below ℃ to obtain a very fine and uniform emulsion.

[0023]

The present invention is further illustrated by the following examples, which do not limit the scope of the claims.
In describing the examples, the trade names of the products are used.
It should be considered that the product may be any product of the same characteristics marketed by another company. The products are: "Miglyol 812" (registered trademark; Dynamit Nob)
el), Sweden]: C fractionated from coconut oil
Triglycerides of _{8 ~C 10} fatty acid. "Edenor (Edenor) SbO _5" [registered trademark;
Henkel, Düsseldorf]: A mixture of saturated and unsaturated fatty acids whose main constituent is oleic acid (67%). "Lutrol F68" [registered trademark; B
ASF, Germany]: Poloxamer which is a polyoxyethylene-polyoxypropylene copolymer
er) 188. In all of the formulations below, the two phases are separately sterilized to prepare the emulsion aseptically, or the final product is sterile filtered through a 0.22 μm filter.

Example 1: Miglyol 812 (registered trade name)
Nanoemulsion mark) (Emarujo emissions A) nonionic surfactant (Lutrol F68 (R))
Add 10 g to 500 ml deionized water. To this solution is added 15 ml of oil (Miglyol 812®). It is then passed through an Ultra-Turrax homogenizer (Junk & Kunkel, Staofen, Germany) at 10,000 rpm for 20 minutes to emulsify it and obtain a nanoemulsion with a particle size smaller than 0.5 μm. 0.25 g disodium edetate (stabilizer), 27.4 g sorbitol powder (isoosmotic agent)
And 0.05 g of benzalkonium chloride (preservative) are added to the emulsion.

The concentrations of the resulting solutions are as follows: Lutrol F68®: 2.00% (weight / volume) Miglyol 812®: 3.00% (vol / vol) Disodium Edible Tate: 0.05% (weight / volume) Sorbitol powder: 5.48% (weight / volume) Benzalkonium chloride: 0.01% (weight / volume) Add deionized water to bring the total volume to 100 ml. Zetasizer 3 [Malvern Instruments]
Company, UK], the average particle size of the oil droplets was 230 n.
m and the polydispersity was 0.220.

Example 2: Miglyol 812 (registered trade name)
Except adding 20g nanoemulsion (Emarujo emissions B) Lutrol F68 in mark) instead of adding 10g was conducted according to the method described in Example 1. The resulting concentrations are as follows: Lutrol F68®: 4.00% (weight / volume) Miglyol 812®: 3.00% (vol / vol) Disodium Ethate: 0.05 % (Weight / volume) Sorbitol powder: 5.48% (weight / volume) Benzalkonium chloride: 0.01% (weight / volume) Add deionized water to make a total volume of 100 ml. As a result of measurement with Zetasizer 3, the average particle size of the oil droplets was 237 nm, and the polydispersity was 0.241.

Example 3: Carteolol base 0.2
8% of the nanoemulsion (emulsion C) Lutrol F68® is added to 190 ml of deionized water and the whole solution is filtered through 0.22 μm (aqueous phase). Separately weigh 0.44 g of carteolol base and add 0.2 g of edenol SbO ₅ and 2.0 g of Miglyol 812®. Heat gently until a carteolol-based solution is obtained (oil phase). The oil phase is added to the aqueous phase while stirring with a magnetic stirrer, which is then passed through an Ultra-Turrax homogenizer at 10,000 rpm for 10 minutes until a nanoemulsion with a particle size smaller than 0.5 μm is obtained. The nanoemulsion thus obtained has a basic pH,
Neutralize to pH = 7.4 with 0.1N hydrochloric acid solution. To render the emulsion isotonic, 10.14 g of apyrogenic mannitol are added thereto, followed by 2 ml of a 1% (weight / volume) benzalkonium chloride solution. Finally, add deionized water and add 20
Finish with 0 ml.

The final concentrations are as follows: Lutrol F68®: 4.00% (weight / volume) Miglyol 812®: 1.00% (weight / volume) Edenol SbO ₅ : 0.0 10% (weight / volume) Carteolol base: 0.22% (weight / volume) Apyrogenic mannitol 5.07% (weight / volume) Benzalkonium chloride: 0.01% (weight / volume) Deionized water To make a total volume of 100 ml. As a result of measurement with Zetasizer 3, the average particle size of the oil droplets was 272 nm and the polydispersity was 0.273.

Example 4: 0.1% of indomethacin
No. Emulsion (Emulsion D) 1.66 g of Lutrol F68 (registered trademark) was added to 0.22 µ
Dissolve in 190 ml of deionized water filtered through an aqueous solution (aqueous phase). 0.05 g of indomethacin and Miglyol 8
0.5 g of 12® are dissolved in 50 ml of acetone (organic phase). The organic phase is added to the aqueous phase while stirring with a magnetic stirrer at 500 rpm. The resulting dispersion is evaporated on a rotary evaporator under reduced pressure and at a temperature below 35 ° C. until all of the acetone and some of the water have been removed and the final volume is 40 ml. 2.53g to make this osmotic pressure
Add aprogenic mannitol, 1% as preservative
0.50 ml of (weight / volume) benzalkonium chloride are added. Thereto, 0.0025 g of crystallized monopotassium phosphate and 0.1128 g of crystallized disodium phosphate dodecahydrate were added and dissolved to prepare a buffer.
Adjust H to 7. Add 0.0275 g of disodium edetate as a stabilizer and add
Finish to 0 ml.

The concentrations of the obtained are as follows: Lutrol F68®: 3.320% (weight / volume) Miglyol 812®: 1.000% (weight / volume) Indomethacin: 0.1% 100% (weight / volume) Aprogenic mannitol: 5.070% (weight / volume) Crystalline monopotassium phosphate: 0.005% (weight / volume) Crystalline disodium phosphate dodecahydrate: 0.221% ( (Weight / volume) Disodium edetate: 0.055% (weight / volume) Benzalkonium chloride: 0.010% (weight / volume) Add deionized water to bring the total volume to 100 ml. As a result of measurement with Zetasizer 3, the average particle size of the oil droplets was 280 nm and the polydispersity was 0.250.

Example 5: Miglyol 812 (registered trade name)
Lutrol nanoemulsion gel (of 0.10g of deionized water Emma Rujon E) 15 ml of mark) F68
® is added and the whole solution is filtered through 0.22 μm. While stirring with a magnetic stirrer, add 0.
20 ml of Miglyol 812® are added.
Next, the resulting dispersion is emulsified in an Ultra-Turrax homogenizer at 10,000 rpm for 15 minutes to obtain a nanoemulsion having a particle size of less than 0.5 μm. To this emulsion are added 1.014 g of aprogenic mannitol and 0.2 ml of a 1% (weight / volume) solution of benzalkonium chloride. To finish the formulation, use the previously prepared Carbol 940 0.4%.
Add 5 g of 6% gel. Stir with a glass rod until a gel of the desired viscosity is obtained.

The concentrations of the preparations are as follows: Lutrol F68®: 0.50% (weight / volume) Miglyol 812®: 1.00% (weight / volume) Aprogenic mannitol : 5.07% (weight / volume) Benzalkonium chloride: 0.01% (weight / volume) Carbopol 940: 0.15% (weight / volume) Add deionized water to bring the total volume to 100 ml. . As a result of measurement with Zetasizer 3, the average particle size of the oil droplets was 278 nm and the polydispersity was 0.259.

Example 6: Miglyol 812 (registered trade name)
Nanoemulsion (emulsion F) 4.00 g Lutrol F6 in 190 ml deionized water
8® is added and the whole solution is filtered through 0.22 μm. Once dissolved, the resulting aqueous phase is placed in a 70 ° C. water bath. Once at that temperature, 6.00 ml of Miglyol 812® are added. The resulting dispersion is mixed until an emulsion smaller than 0.5 μm in particle size is obtained.
Run in an Ultra-Turrax homogenizer at 000 rpm for 15 minutes. Then 10.96 g sorbitol powder, 0.10 g disodium edetate and 1%
1 ml of benzalkonium chloride (weight / volume) concentration is added. Finally, make up the volume to 200 ml with deionized water.

The final concentrations are as follows: Lutrol F68®: 2,000% (weight / volume) Miglyol 812®: 3.000% (weight / volume) Sorbitol powder: 5.480 % (Weight / volume) Disodium edetate: 0.050% (weight / volume) Benzalkonium chloride: 0.005% (weight / volume) Add deionized water to bring the total volume to 100 ml. As a result of measurement with Zetasizer 3, the average particle size of the oil droplets was 224.3 n.
m and the polydispersity was 0.175.

Example 7: Miglyol 812 (registered trade name)
Lutrol 0.5g of the nanoemulsion (Emarujo emission G) deionized water 90ml of target) F68
® is added and the whole solution is filtered through 0.22 μm. Once dissolved, the resulting aqueous phase is placed in a 70 ° C. water bath. At that temperature, 1.00 ml of Miglyol 812® is added. The dispersion obtained is reduced to 10 by 10 until an emulsion smaller than 0.5 μm in particle size is obtained.
Run in an Ultra-Turrax homogenizer at 000 rpm for 10 minutes. Then 5.48 g of sorbitol powder and 1 ml of 1% (weight / volume) concentration of benzalkonium chloride
Add. The volume is then made up to 10 ml with deionized water.

The final concentrations are as follows: Lutrol F68®: 0.50% (weight / volume) Miglyol 812®: 1.00% (weight / volume) Sorbitol powder: 5.48 % (Weight / volume) Benzalkonium chloride: 0.01% (weight / volume) Add deionized water to bring the total volume to 100 ml.

Stability Evaluation The stability of Emulsion A and Emulsion B kept at different temperatures was tracked. The controls are run for different times,
The results for the average particle size of the oil droplets are shown in FIGS. 1 and 2, and the results for the polydispersity are shown in FIGS. The pH results are shown in FIGS. No significant changes were observed in any of the three parameters during the entire period of the evaluation. In nanoemulsion C containing carteolol base, in addition to the above parameters, the active substance content was also confirmed. The average particle size results are shown in FIG. 7, the polydispersity results are shown in FIG. 8, and the pH results are shown in FIG. The result of the active substance content is shown in FIG. No significant changes in physico-chemical parameters have been observed. As seen in FIG. 10, the freezing temperature,
There is no change in carteolol base concentration during storage at room temperature of 35 ° C and 5-45 ° C (temperature alternation).

Study of Acute Ocular Tolerance of Rabbit Emulsion B and E were repeatedly infused with 50 μl of Emulsion B and E every 20 minutes for 20 minutes in rabbits of New Zealand albino. Was evaluated. Tolerable concentrations for the eyes were evaluated 24 hours after the application was completed, by blinking, redness, edema, exudate and damage in the iris and cornea. The results show that emulsions B and D have a normal eye irritation index.

The "in vivo" corneal permeation penetration Emulsion of indomethacin contained at a concentration of 0.1% (weight / volume) in nanoemulsion D was evaluated and it was estimated to be 0.1% (weight / volume) in aqueous solution. Compared to the penetration of indomethacin contained in the concentration. For this purpose, rabbit corneas were removed from Ziesz-Noguer.
a) and the like.
original Penetration Chambe
r) LC-100 "[(3) Dies-Noguera A., Igual A. and Guzma L.
nL.), “Design of Ann“ In Vitro ”
・ System for Pharmacokinetic Studies (Design of an “in vitro”)
system for pharmacokinetic
Laboratories Kushi, Labs. CUSI, SA, El Masnous, Barcelona (Bar)
celona), Catalonia,
Spain, 7th edition, International Congress of the Eye Research (International C)
ong. of Eye Research, Nagoya, Japan (1986)] (FIG. 11). The corneal coating surface was exposed to the test product for 3 hours. 200 μl of an aliquot of artificial body fluid (AAH) was added to 15, 30, 6
Withdrawn from the rear of the chamber after 0, 120, 150 and 180 minutes. The Wach sample was immediately replaced with an equal amount of AAH.

The indomethacin content of the samples was measured by HPLC immediately after extraction and at 250 nm, and the values obtained were used to calculate the osmotic coefficient (P, in cm / s). INDO: 0.1% (weight / volume) indomethacin solution NAND: nanoemulsion D The results are shown graphically in FIG.

The permeability coefficient is calculated by the equation of Glass and Robinson [(4) Grass G.M.
And Robinson J. Earl (Robinson
J.R.), Journal of Pharmaceutical Sciences (Journal of Pharm)
Aceutical Sciences), Vol. 77,
No. 1, pp. 3-14 (1988)]

(Equation 1) In the formula, Δt / Δc: slope of the linear portion of the graph 3: volume in ml on the back of the chamber 60: conversion factor from minute to second A: corneal surface exposed to the product (0.721 cm ² ) Co: It was calculated using the theoretical concentration of the test product. Latency was obtained by extrapolating the linear portion of the graph. As can be seen from FIG. 12, indomethacin in the nanoemulsion penetrates earlier and more often than when in solution.

“In vivo” Pharmacokinetic Studies Pigmented rabbits
[Fouver de Bourgogne (Fauver de
Bourgonge)] and nanoemulsion D and 0.1
% (Weight / volume) aqueous indomethacin solution
One infusion of 5 μl was performed. Approximately 200 μl of aqueous bodily fluid from the front chamber was infused 15 minutes, 30 minutes, and 1, 2,
Samples were collected at 4, 6, and 8 hours. The aqueous body fluid sample was
Filtered through a 45 μm filter and analyzed by HPLC immediately after collection and at 250 nm. Table 1 shows the obtained results.

[0043]

[Table 1]

The results are shown as a graph in FIG. FIG.
As can be seen from FIG. 3, the experimental data indicate that indomethacin in Nanoemulsion D has a significant increase in penetration compared to aqueous solutions. Similarly, the expression of indomethacin in the body fluid also exceeded the detection limit by 2 hours in the case of nanoemulsion D, as compared with the case of the aqueous solution.

Preservation Efficacy Studies Bacteria were incubated in tryptose-soybean at 34 ° C. for 1 hour.
The cells were cultured for 8 hours. Candida albicans and Alpergillus niger were cultured in a Sabouraud culture medium in a 22 ° C. oven for 48 hours and 7 days, respectively. A suspension of about 1 × 10 ⁸ ufc / ml was prepared from the culture medium. 20 ml of each nanoemulsion (A, F and G)
200 μl of each microorganism suspension was added to the tube containing.
The microbial ufc contained in 1 ml of the nanoemulsion at 0, 6, 24 hours and 7, 14 and 28 days was counted. Furthermore, the microorganisms in physiological saline were counted as inoculated controls, and the sterilized control in the uninoculated medium was counted. For each ml of each nanoemulsion, and for each test time, add 0.1 to neutralize the preservative.
A series of dilutions (1/19) were prepared diluted in Leethan broth containing 5% (weight / volume) Tween 80. 1 ml of each dilution
Was melted at 45 ° C. and cultured in triplicate in 20 ml of tryptose agar soybeans supplemented with 0.5% (w / v) Tween 80.

Nanoemulsions A and F are European
Pharmacopeia 1993 (Euroean Pha
rmacopoeia 1993) (Reference B) and British Pharmacopeia 1993 (Bri
tissue Pharmacopoeia 1993) (Criterion B); Pharmacopee Francaisé 1989 (Farmacopee Francaisé 198).
9) and USP, Vol. 22, 19
Complies with 90-year rules. Nanoemulsion G with less surfactant is also European pharmacopeia 1
993 (criteria A) and British Pharmacopeia 1993 (criteria A). Thus, the nanoemulsions of the present invention have much lower preservative concentrations than are used in other oil-in-water emulsions and meet most important Pharmacopoeia criteria. The latter other oil-in-water emulsions do not meet the pharmacopeia criteria in terms of preservative effectiveness when using the preservative concentrations used in the formulations of the present invention. The results are shown in Tables 2, 3 and 4.

[0047]

[Table 2]

[0048]

[Table 3]

[0049]

[Table 4]

[0050]

[Brief description of the drawings]

FIG. 1 is a graph showing the change over time of the stability of Emulsion A, expressed as the average oil droplet diameter in nm.

FIG. 2 is a graph showing the change over time of the stability of emulsion B, expressed as the average oil droplet diameter in nm.

FIG. 3 is a graph showing the change over time of the polydispersity of Emulsion A.

FIG. 4 is a graph showing the change over time of the polydispersity of emulsion B.

FIG. 5 is a graph showing a time change of the pH of the emulsion A.

FIG. 6 is a graph showing the time change of the pH of emulsion B.

FIG. 7 is a graph showing the change over time of the stability of Emulsion C, expressed as the average oil droplet diameter in nm.

FIG. 8 is a graph showing the change over time of the polydispersity of Emulsion C.

FIG. 9 is a graph showing the time change of the pH of emulsion C.

FIG. 10 is a graph showing the change over time in the amount of active substance released in Emulsion C as a percentage of the initial theoretical content.

FIG. 11 is a diagram of a corneal permeation device (model LC-100) used in the “in vitro” test.

FIG. 12 is a graph depicting the permeation of indomethacin through the cornea in Nanoemulsion D, showing the concentration of indomethacin over time.

FIG. 13 “In vivo” for Nanoemulsion D
5 is a graph showing the pharmacokinetics of the indomethacin in the rabbit colored aqueous body fluid over time.

[Explanation of symbols]

1: corneal permeation and permeation chamber 2: constant temperature water bath (37 ° C.) _3. Carbon generation tank (95% O ₂ and 5% CO ₂ ) 4. Omniscribe (universal record) D-5000 recorder 5. Platinum-calomel electrode Agarose: KCl bridge Amplifier 8. Peristaltic pump 9. Magnetic stirrer 10. Artificial tear solution / test product

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁶ Identification code FI A61K 31/405 A61K 31/405 31/47 31/47 31/52 31/52 31/54 31/54 31/57 31/57 38/00 37/02 (72) Inventor Anna Cole Dux Spain 08012 Barcelona, Gran de Gracia 47th Quart Primera (72) Inventor Nuria Carreras Perdigell Spain 08140 Caldes de Montbuy (Barcelona) ), Amadeo Vivez No. 6 (Urbano Els Saurons) (56) References European Patent Application Publication 480690 (EP, A1) (58) Fields investigated (Int. Cl. ⁶ , DB name) A61K 9 / 107 A61K 31/135 A61K 31/165 A61K 31/19 A61K 31/405 A61K 31/47 A61K 31/52 A61K 31/54 A61K 31/57 A61K 38/00

Claims (17)

(57) [Claims] An oil-in-water nanoemulsion useful as an ophthalmic vehicle, comprising oil droplets, obtained by preparing an emulsion of an oil phase in an aqueous phase, comprising: 0.1 to 10 % (Weight / volume) oil; 0.1-10% (weight / volume) nonionic surfactant; less than 0.01% (weight / volume) benzalkonium chloride as preservative; 0.01 to 5% (weight / volume) of a medicament, a pharmaceutical precursor or a biologically active substance; an optional component, and also various proportions of isotonic agents, viscosity modifiers, stabilizers And at least one component selected from buffers and antioxidants; having a pH of 5 to 8 and an osmotic pressure of 250 to 400 mOsm.
The surfactant is from the aqueous phase and is miscible with water; its transmittance at 520 nm is less than 70%; the drug, drug precursor or biologically active substance is an oil droplet A nanoemulsion characterized by being contained therein. 2. The nanoemulsion according to claim 1, wherein the oil is a fractionated fatty acid triglyceride of coconut oil, a mixture of fatty acids or a propylene glycol ester of saturated vegetable fatty acids. 3. The nanoemulsion according to claim 1, wherein the nonionic surfactant is a polyoxyethylene-polyoxypropylene copolymer. 4. The method according to claim 1, wherein the nonionic surfactant is poloxamer (p
4. A nanoemulsion according to claim 3, which is an oxamer. 5. The nanoemulsion according to claim 1, comprising a drug selected from anti-glaucoma drugs, antibiotics, antibiotic allergic drugs, antiviral drugs and anti-inflammatory drugs. 6. An anti-glaucoma drug comprising carteolol-based (c)
arteolbase, betaxolol (bet)
The nanoemulsion according to claim 5, wherein the nanoemulsion is selected from axolol, atenolol, and timolol base. 7. The method of claim 1, wherein the anti-inflammatory drug is indomethacin (indom).
ethacin), piroxicam (pyroxyca)
m), dichlorofenac acid (dichlofenac)
acid), ibuprofen
n), dexamethasone
6. The nanoemulsion according to claim 5, wherein the nanoemulsion is selected from the group consisting of clobetasone and clobetasone. 8. The method according to claim 8, wherein the drug is cyclosporin.
The nanoemulsion according to claim 5, which is (porin) A. 9. The method according to claim 9, wherein the drug is acyclovir.
ovir; 2-amino-1,9-dihydro-9-[(2-hydroxy-ethoxy) methyl] -6H-purin-6-one). 10. The method according to claim 10, wherein the medicine is acid chromoglygate (acid).
The nanoemulsion according to claim 5, which is a chromolygate. 11. The nanoemulsion of claim 1, wherein the isotonic agent is selected from mannitol, glycerol, sorbitol and glucose. 12. The nanoemulsion according to claim 1, wherein the viscosity modifier is selected from hydroxyopropylmethylcellulose, a polyacrylic acid derivative, and sodium carboxymethylcellulose. 13. The nanoemulsion of claim 1, wherein the stabilizer is selected from sodium ethylenediaminetetraacetate and citric acid. 14. The nanoemulsion of claim 1, wherein the buffer is selected from sodium phosphate, potassium phosphate, sodium citrate, sodium carbonate and sodium bicarbonate. 15. The method according to claim 1, wherein the medicament, the drug precursor or the biologically active substance is partially or completely dissolved in the oil. Nanoemulsion. 16. 0.005% of benzalkonium chloride
The nanoemulsion according to claim 1, which is found at a concentration equal to or greater than (weight / volume). 17. Preparing an aqueous phase containing a water-soluble nonionic surfactant and optionally further components selected from iso-osmotic agents, preservatives, buffers and viscosity modifiers; oils and pharmaceuticals , An active ingredient selected from the group of pharmaceutical precursors and biologically active substances and partially or completely dissolved in oil or a mixture of these active ingredients, and an oil phase containing an antioxidant as an optional ingredient. Preparing; dissolving the oil phase in an organic solvent that is miscible with water and having a dielectric constant greater than 15 to prepare the organic phase; adding the organic phase to the aqueous phase under gentle stirring Removing an organic solvent and some water at a temperature of 35 ° C. or less under reduced pressure to obtain a very fine and uniform emulsion containing oil droplets having an average particle diameter of less than 500 nm. Nanoemers described in Manufacturing method of John.